AZD 4017

AZD 4017 displays no detectable activity against glucocorticoid and mineralocorticoid receptors, but demonstrates remarkable selectivity against related enzymes 11-βHSD2, 17β-HSD1, and 17β-HSD3 (all IC50>30 μM). With the exception of cynomolgus monkeys (IC50=0.029 μM), AZD 4017 has markedly reduced activity in various species, despite its high potency against the human version of 11β-HSD1. Furthermore, as adipose tissue is believed to be an important target organ, suppression of 11β-HSD1 activity was assessed in isolated human adipocytes from individuals without diabetes. There is considerable certainty that AZD 4017 is not restricted by adipose tissue because of its acidic nature because it was demonstrated to be a strong inhibitor of this important target tissue (IC50=0.002 μM), which is in good accord with the potency of the enzyme [1].

AZD 4017 对糖皮质激素和盐皮质激素受体没有表现出可检测的活性，但对相关酶 11-βHSD2、17β-HSD1 和 17β-HSD3 显示出显着的选择性（所有 IC50>30 μM）。除食蟹猴外（IC50=0.029 μM），AZD 4017 在多种物种中的活性均显着降低，尽管它对人类版本的 11β-HSD1 具有高效能。此外，由于脂肪组织被认为是重要的靶器官，因此在来自非糖尿病个体的分离的人类脂肪细胞中评估了 11β-HSD1 活性的抑制。可以相当肯定的是，AZD 4017 因其酸性性质而不受脂肪组织的限制，因为它被证明是这一重要靶组织的强抑制剂 (IC50=0.002 μM)，这与酶的效力非常一致[1]。

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AZD4017 能有效抑制人11β-HSD1酶活性，IC50为7 nM。
在从非糖尿病志愿者分离的人脂肪细胞中，AZD4017 抑制可的松向皮质醇的转化，IC50为2 nM，证实了其在关键靶组织中的活性。
该化合物对五种主要的细胞色素P450酶（CYP1A2、CYP2C9、CYP2C19、CYP2D6和CYP3A4）无显著抑制（在10 μM浓度下抑制率<25%）。
在两株Ames试验中无致突变性，在小鼠淋巴瘤试验中为阴性。
在hERG IonWorks试验中，IC50 > 100 μM。
在MDCK细胞渗透性试验中表现出良好的渗透性，表观渗透率（Papp A-B）为29 x 10^-6 cm/s，外排比为0.8。
晶体结构揭示了其结合模式：酰胺羰基与Tyr183相互作用，羧酸盐与Leu217的骨架NH形成氢键，吡啶环位于疏水残基Leu217和Ala172之间。 [1]

由于 AZD 4017 对小鼠酶的功效较低，仅进行了一些临床前药效学评估。增加剂量进一步在 1500 mg/kg 时产生约 70% 抑制的最大效果，这与小鼠中的 10×IC50 相当。这表明 AZD 4017 在此模型中以剂量依赖性方式抑制 11β-HSD1 [1]。

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在使用经口服给予AZD4017的瘦型小鼠附睾脂肪组织进行的离体试验中，观察到了剂量依赖性的11β-HSD1活性抑制。
给药后1小时，在50 mg/kg的剂量下达到50%抑制，对应的血浆暴露量相当于小鼠酶的IC50。
在1500 mg/kg（相当于小鼠酶IC50的10倍）的剂量下，观察到约70%的最大抑制效应。 [1]

使用皮质醇竞争性均相时间分辨荧光（HTRF）法测定化合物对11β-HSD1的抑制活性。
将含有可的松、葡萄糖-6-磷酸、NADPH、葡萄糖-6-磷酸脱氢酶、EDTA、磷酸钾缓冲液（pH 7.5）、重组人（或其他物种）11β-HSD1酶和待测化合物的反应体系加入黑色384孔板中。
将板在37°C下孵育25分钟，使可的松酶促转化为皮质醇。
通过加入含有皮质醇-XL665示踪剂和酸的溶液终止反应。
随后加入抗皮质醇穴状化合物抗体，在室温下孵育2小时后，使用酶标仪在320 nm激发下测量665 nm和620 nm的荧光发射。
计算665 nm与620 nm的荧光比值，并用于确定IC50值。 [1]

评估了化合物在分离的人脂肪细胞中对11β-HSD1的抑制。
从非糖尿病志愿者获取皮下脂肪组织，并通过胶原酶消化分离脂肪细胞。
将分离的脂肪细胞在含有^3H-可的松和待测化合物的培养基中，于37°C、5% CO2条件下孵育6小时。
孵育后，收集培养基，用乙酸乙酯萃取放射性标记的类固醇。
将萃取样品干燥、复溶，并使用C18色谱柱，以甲醇/水为流动相，通过反相高效液相色谱（HPLC）进行分析。
使用流式闪烁分析仪测量洗脱出的对应于可的松和皮质醇的放射性，以定量酶促转化及其抑制程度。 [1]

For the ex vivo pharmacodynamic study in mice, male C57BL/6 mice on a chow diet were dosed by oral gavage with AZD4017 suspended in a hydroxypropyl methylcellulose (HPMC)/Tween formulation.
At 1 hour post-dose, animals were euthanized, blood was collected via cardiac puncture for compound level analysis, and epididymal fat pads were removed and snap-frozen.
The adipose tissue was cut into small pieces and incubated in medium containing ^3H-cortisone for 1 hour at 37°C.
Following incubation, the medium was processed for steroid extraction and HPLC analysis as described in the cell assay to measure residual 11β-HSD1 activity. [1]
For pharmacokinetic studies in mouse, rat, and dog, AZD4017 was administered at doses of 1-3 mg/kg, either as a solution in DMSO/hydroxy-β-cyclodextrin or as a suspension in HPMC/Tween. [1]

AZD4017 exhibited high oral bioavailability: >100% in mouse and rat (attributed to enterohepatic recirculation of an acyl glucuronide metabolite), and 60% in dog.
Plasma clearance (Clp) was moderate: 16 mL/min/kg in mouse, 9 mL/min/kg in rat, and 7 mL/min/kg in dog.
Volume of distribution at steady state (Vd_ss) was: 4.2 L/kg in mouse, 2.6 L/kg in rat, and 0.9 L/kg in dog.
Half-life (t1/2) ranged from 3.2 to 4.7 hours in mouse and rat, and was 4.9 to 6.2 hours in dog.
Plasma protein binding (PPB) was low across species, with free fractions of: mouse 2.9%, rat 1.8%, dog 2.3%, and human 0.8%.
Aqueous solubility at pH 7.4 was 144 μM.
The major route of metabolism in rat, dog, and human hepatocytes was conversion to the acyl glucuronide conjugate.
Secondary peaks in PK profiles of mouse and dog suggested enterohepatic recirculation. [1]

A radiolabeled covalent binding study in male rats dosed orally with 20 mg/kg of AZD4017 showed no significant covalent binding to plasma or liver proteins (<25 pmol/mg protein at 2 hours, undetectable at 24 hours).
No significant induction of cytochrome P450 enzymes was observed in rats after 1 month of administration.
The compound was assessed in a guinea pig monophasic action potential (MAP) assay with no changes observed in any parameters.
It showed a clean selectivity profile against a broad panel of off-target enzymes and receptors. [1]

[1]. Discovery of a potent, selective, and orally bioavailable acidic 11β-hydroxysteroid dehydrogenase type 1 (11β-HSD1) inhibitor: discovery of 2-[(3S)-1-[5-(cyclohexylcarbamoyl)-6-propylsulfanylpyridin-2-yl]-3-piperidyl]acetic acid (AZD4017). J Med Chem. 2012 Jun 28;55(12):5951-64.

AZD-4017 is under investigation in clinical trial NCT01173471 (A Phase IIa Study to Assess the Tolerability, Safety, and Efficacy of AZD4017 for Raised Intra-ocular Pressure).
11beta-hydroxysteroid Dehydrogenase Type 1 Inhibitor AZD4017 is a selective, orally bioavailable inhibitor of the enzyme 11beta-hydroxysteroid dehydrogenase type 1 (11b-HSD1; 11bHSD1; HSD11B1), with potential protective activity. Upon administration, AZD4017 selectively binds to and inhibits the activity of 11b-HSD1. This prevents the conversion of cortisone to the active hormone cortisol that activates glucocorticoid receptors. By blocking cortisol production in metabolic tissues, AZD4017 may inhibit the adverse metabolic effects, such as glucose intolerance, hyperinsulinemia, systolic hypertension, increased adiposity, myoatrophy and dermal atrophy, caused by exogenous administration of glucocorticoids in these tissues. 11bHSD1 is highly expressed in metabolic tissues, such as liver, skeletal muscle, and adipose tissue. It plays a crucial role in regenerating active glucocorticoid from circulating glucocorticoids and regulates the production of cortisol to activate the glucocorticoid receptors. AZD4017 does not inhibit the anti-inflammatory activity of the administered glucocorticoids.

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AZD4017 is an acidic inhibitor discovered from a 2-thioalkylnicotinamide series, optimized to introduce a carboxylic acid group for improved metabolic stability and pharmacokinetics.
Its high ligand lipophilicity efficiency (LLE = 6.0) indicated an optimal balance of potency and lipophilicity.
One stable anhydrous crystalline form was identified, suggesting no significant solid-state development issues.
Based on its potent target inhibition, excellent selectivity, favorable pharmacokinetic profile across species, and clean preliminary safety pharmacology, AZD4017 was selected as a clinical candidate for the treatment of metabolic syndrome. [1]

C22H33N3O3S

419.58100

419.224

C, 62.98; H, 7.93; N, 10.01; O, 11.44; S, 7.64

1024033-43-9

24946280

Light yellow to yellow solid powder

4.793

107.83

2

6

8

29

541

1

O=C(O)C[C@H]1CN(C2=NC(SCCC)=C(C(NC3CCCCC3)=O)C=C2)CCC1

NCDZABJPWMBMIQ-INIZCTEOSA-N

InChI=1S/C22H33N3O3S/c1-2-13-29-22-18(21(28)23-17-8-4-3-5-9-17)10-11-19(24-22)25-12-6-7-16(15-25)14-20(26)27/h10-11,16-17H,2-9,12-15H2,1H3,(H,23,28)(H,26,27)/t16-/m0/s1

2-[(3S)-1-[5-(cyclohexylcarbamoyl)-6-propylsulfanylpyridin-2-yl]piperidin-3-yl]acetic acid

AZD4017; AZD-4017; AZD 4017.

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

DMSO : ~125 mg/mL (~297.92 mM)

配方 1 中的溶解度: ≥ 2.08 mg/mL (4.96 mM) (饱和度未知) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (这些助溶剂从左到右依次添加，逐一添加), 澄清溶液。
例如，若需制备1 mL的工作液，可将100 μL 20.8 mg/mL澄清DMSO储备液加入400 μL PEG300中，混匀；然后向上述溶液中加入50 μL Tween-80，混匀；加入450 μL生理盐水定容至1 mL。
*生理盐水的制备：将 0.9 g 氯化钠溶解在 100 mL ddH₂O中，得到澄清溶液。

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配方 2 中的溶解度: ≥ 2.08 mg/mL (4.96 mM) (饱和度未知) in 10% DMSO + 90% Corn Oil (这些助溶剂从左到右依次添加，逐一添加), 澄清溶液。
例如，若需制备1 mL的工作液，可将 100 μL 20.8 mg/mL 澄清 DMSO 储备液加入到 900 μL 玉米油中并混合均匀。

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请根据您的实验动物和给药方式选择适当的溶解配方/方案：
1、请先配制澄清的储备液(如：用DMSO配置50 或 100 mg/mL母液(储备液));
2、取适量母液，按从左到右的顺序依次添加助溶剂，澄清后再加入下一助溶剂。以 下列配方为例说明 (注意此配方只用于说明，并不一定代表此产品 的实际溶解配方):
10% DMSO → 40% PEG300 → 5% Tween-80 → 45% ddH2O (或 saline);
假设最终工作液的体积为 1 mL, 浓度为5 mg/mL: 取 100 μL 50 mg/mL 的澄清 DMSO 储备液加到 400 μL PEG300 中，混合均匀/澄清；向上述体系中加入50 μL Tween-80，混合均匀/澄清；然后继续加入450 μL ddH2O (或 saline)定容至 1 mL；
3、溶剂前显示的百分比是指该溶剂在最终溶液/工作液中的体积所占比例;
4、 如产品在配制过程中出现沉淀/析出，可通过加热(≤50℃)或超声的方式助溶;
5、为保证最佳实验结果，工作液请现配现用！
6、如不确定怎么将母液配置成体内动物实验的工作液，请查看说明书或联系我们;
7、 以上所有助溶剂都可在 Invivochem.cn网站购买。